Cd27 fitc

Manufactured by BioLegend

CD27-FITC is a fluorochrome-conjugated antibody that binds to the CD27 cell surface antigen. CD27 is a member of the tumor necrosis factor receptor superfamily and is expressed on T cells, B cells, and natural killer cells. The FITC fluorochrome allows for the detection and analysis of CD27-positive cells using flow cytometry.

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Lab products found in correlation

Cd19 apc, by BD (2 mentions) Cd27 apc, by BD (1 mentions) Cd38 apc, by BD (1 mentions) Cd8 pe, by Beckman Coulter (1 mentions) Cd38 pacific blue, by Beckman Coulter (1 mentions) Cd16 cd56 pe, by Beckman Coulter (1 mentions) Cd45ra pe, by Beckman Coulter (1 mentions) Cd27 bv605, by BD (1 mentions) Cd27 percp cy5, by BD (1 mentions) Cd3 apc, by Beckman Coulter (1 mentions) Facscelesta, by BD (1 mentions) Cd4 fitc, by BD (1 mentions) Cd45ra bv605, by BioLegend (1 mentions) Cd8 percp, by BioLegend (1 mentions) Cd20 percp, by BioLegend (1 mentions) Fixable viability stain 510, by BD (1 mentions) Cd14 bv650, by BioLegend (1 mentions) Cd38 bv421, by BioLegend (1 mentions) Hla dr bv605, by BioLegend (1 mentions) Cd16 af700, by BioLegend (1 mentions)

Spelling variants of this product

Anti-CD27-FITC (4 citations) FITC-conjugated CD27 (2 citations) FITC-conjugated anti-CD27 (2 citations)

6 protocols using cd27 fitc

Multiparametric Flow Cytometry Analysis

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For reference, see Stepensky et al. (2013) (link). Additional antibodies used in the study were TCR αβ FITC, CD3 APC, CD16/CD56 PE, CD8 AF700, CD45RA PE, CD38 Pacific Blue, CD20 PerCp, IgG APC, kappa FITC, lambda PE, CD21 FITC, CD8 PE, and TCR γδ PE (Beckman Coulter GmbH); CD45RA APC-Cy7 and IL-2 PerCpCy5.5 (BioLegend Inc.); CD27 FITC, CD27 APC, CD27 BV605, CD27 PerCpCy5.5, CD38 APC, CD19 APC, IgG PE, S6(pS235/236) APC (BD Biosciences), and IgM AF488 (Jackson); and BclXL AF488 (Cell Signaling Technologies).

Keller B., Shoukier M., Schulz K., Bhatt A., Heine I., Strohmeier V., Speckmann C., Engels N., Warnatz K, & Wienands J. (2018). Germline deletion of CIN85 in humans with X chromosome–linked antibody deficiency. The Journal of Experimental Medicine, 215(5), 1327-1336.

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Multiparametric Immune Profiling of PBMC

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Cryopreserved PBMCs were thawed and left for 1 h at 37 °C in the CO₂ incubator. Subsequently the cells were collected and stained with BD Horizon™ Fixable Viability Stain 510 to identify live cells, as per manufacture protocol. The cells were then washed, and surface stained for IL-21R PE expression on, (i) CD4, CD8 and Tfh (CD4/CD45RA -/CXCR5⁺/PD1⁺); (ii) B cell CD20, B1(CD20⁺CD27⁺CD43⁺), Plasma blast (CD20⁺CD38⁺) and iii) monocytes CD14⁺HLADR⁺.
After staining, the cells were washed and fixed using 2% PFA. Acquisition was done on BD FACSCelesta (Becton-Dickenson, San Jose, CA). Forward and side scatters and singlets were used to gate and exclude cellular debris. The flow cytometry results were analyzed using FlowJo™ v10.8 Software (BD Life Sciences, Ashland, OR). The details of the antibodies used are as follows: Fixable Viability stain510, CD4 FITC (clone: RPA-T4) from BD Bioscience (San Jose, CA), IL-21R PE (clone: 2G1-K12), CD8 PerCP (clone: SK1), PD1 APC (clone: EH12.2H7), CXCR5 BV421 (clone: J252D4), CD45RA BV605 (clone: HI100), CD38 BV421 (clone: HB-7), CD16 AF700 (clone: B73.1), CD14 BV650 (clone: M5E2), HLADR BV605 (clone: L243), CD20 PerCP (clone: 2H7), CD27 FITC (clone: M-T271), CD43 APC (clone: CD43-10G7) from BioLegend (San Diego, CA).

Agrawal S., Baulch J.E., Madan S., Salah S., Cheeks S.N., Krattli RP J.r., Subramanian V.S., Acharya M.M, & Agrawal A. (2022). Impact of IL-21-associated peripheral and brain crosstalk on the Alzheimer’s disease neuropathology. Cellular and Molecular Life Sciences, 79(6), 331.

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PBMC Isolation and Immunophenotyping

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Peripheral blood mononuclear cells (PBMCs) were isolated from blood samples with an EDTA tube by density gradient centrifugation using Ficoll–Paque, then resuspended in a freezing solution and stored in liquid nitrogen. Frozen PBMCs were thawed and washed, then stained with fluorescently labeled monoclonal antibodies [anti-human CD3-FITC, CD16/CD56-PE, CD45-PerCP, CD19-APC (Beckton Dickens (BD), MultitestTM), CD19-APC, CD27-FITC, CD24-PerCP, CD38-Alexa Fluor 700, IgD-APC/Cy7, and CD138-PE (Biolegend)] and, then six color immunofluorescence staining was utilized (BD FACS Aria II, Becton Dickinson, Franklin Lakes, NJ). Data were analyzed using Cell Quest (BD) and FlowJo7.6.5 software.

Ozdag Acarli A.N., Tuzun E., Sanli E., Koral G., Akbayir E., Cakar A., Sirin N.G., Soysal A., Aysal F., Durmus H., Parman Y, & Yilmaz V. (2023). Disease activity in chronic inflammatory demyelinating polyneuropathy: association between circulating B-cell subsets, cytokine levels, and clinical outcomes. Clinical and Experimental Immunology, 215(1), 65-78.

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Murine Hematopoietic Stem Cell Analysis

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For bone marrow, cells were stained with antibodies to (hybridoma names and vendors in parentheses): Kit-APC/Cy7 (2B8, Becton Dickinson), CD27-FITC (LG.3A10, Biolegend), CD201-APC (RCR-16, Biolegend), Sca-1 (D7, BV605, BD; and E13-161-7, Pacific Blue, Biolegend), CD150-Biotin (TC15-12F12.2; with streptavidin-Brilliant Violet 421, BD), CD4 (GK1.5), CD8 (53-6.7), CD3ε (145-2C11), Ter119 (TER-119), B220 (RA3-6B2), Gr1 (RB6-8C5), Mac-1 (M1/70) were used. Antibodies to CD4, CD8, CD3ε, Ter119, B220, Gr-1 and Mac-1 were purchased as conjugates to PE/Cy5 (Biolegend). For peripheral blood, cells were stained with antibodies to: CD3ε-Pacific Blue (17A2, Biolegend), CD3ε-PE (145-2C11, Biolegend) CD45-PECy5 (30-F11, Biolegend), Mac-1-PECy7 (M1/70, Biolegend), Gr1-Alexa Fluor 680 (in house), B220-Pacific Blue (RA3-6B2, Biolegend), B220-FITC (in house). Cells were stained on ice in staining buffer for 1 hour, washed, and resuspended in staining buffer with propidium iodide (0.25ug/ml). Analysis and sorting of stained cells were performed on an LSRII (BD Biosciences) or on a FACSAria (BD Biosciences) at the Joslin Diabetes Center Flow Core.

Vazquez S.E., Inlay M.A, & Serwold T. (2015). CD201 and CD27 identify hematopoietic stem and progenitor cells across multiple murine strains independently of Kit and Sca-1. Experimental hematology, 43(7), 578-585.

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Detailed Immunophenotyping Assay Protocol

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Chemicals and reagents were purchased from Sigma Aldrich (St. Louis, MO). Anti–mouse flurochrome labeled antibodies (Abs): CD4-PB, CD8-APC-Cy7, CD62L-APC, CD44-PerCP-Cy5.5, CD127-PE, KLRG1-PE, IFN-γ-PECy7, TNFα-PerCPCy5.5, IL-17-PerCPCy5.5 and Abs for ELISA were procured from BD Pharmingen (San Diego, CA); CD27-FITC, CD43-PE, CXCR3-FITC and CCR6-APC from Biolegend (San Diego, CA) or otherwise mentioned. RPMI-1640 and FBS were purchased from GIBCO (Grand Island, NY). For culturing of cells, tissue culture grade plastic-ware was purchased from BD Biosciences (Bedford, MA).

Rai P.K., Chodisetti S.B., Maurya S.K., Nadeem S., Zeng W., Janmeja A.K., Jackson D.C, & Agrewala J.N. (2018). A lipidated bi-epitope vaccine comprising of MHC-I and MHC-II binder peptides elicits protective CD4 T cell and CD8 T cell immunity against Mycobacterium tuberculosis. Journal of Translational Medicine, 16, 279.

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Comprehensive Immune Cell Profiling

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PBMCs were isolated from whole blood by Ficoll-Paque Plus (GE Healthcare) gradient separation. Fc Receptor Binding Inhibitor (eBioscience) was added to the isolated PBMCs. They were stained with the fluorescent-labeled antibodies for 20 min. The following antibodies were purchased from BioLegend: CD3-PerCP/Cy5.5 (UCHT1), CD3-PE/Cy7 (UCHT1), CD27-FITC (O323), CD38-PE/Cy7 (HIT2), CD19-APC/Cy7 (HIB19), CD45RA-APC/Cy7 (HI100), CD16-PerCP/Cy5.5 (3G8), CD56-APC/Cy7 (HCD56), CCR7-PerCP/Cy5.5 (G043H7), CD14-FITC (M5E2), and CD25-BV421 (BC96). The following antibodies were purchased from BD: CD24-PE (ML5), IgD-BV421 (IA6-2), CXCR5-AF488 (RF8B2), CCR6-PE (11A9), CXCR3-BV421 (1C6/CXCR3), CD4-V500 (RPA-T4) and CD19-V500 (HIB19). CD25-PE/Cy7 (BC96), HLADR-PE (L243), and CD127-PE/Cy7 (eBioRDR5) were purchased from eBioscience, and LAG3-PE (FAB2319P) was purchased from R&D. We classified CD4⁺ T cells, B cells, natural killer (NK) cells and monocytes based on the Human Immunology Project classification (20 (link)). Cell subset definitions of surface markers are shown in Table 1, and the gating strategy is shown in Figure 1. Flow cytometric analyses were performed by 8-color MoFlo XDP (Beckman Coulter). Flow cytometric data were analyzed using FlowJo software (Tree Star).

Kato R., Sumitomo S., Tsuchida Y., Tsuchiya H., Nakachi S., Sakurai K., Hanata N., Nagafuchi Y., Kubo K., Tateishi S., Kanda H., Okamura T., Yamamoto K, & Fujio K. (2019). CD4+CD25+LAG3+ T Cells With a Feature of Th17 Cells Associated With Systemic Lupus Erythematosus Disease Activity. Frontiers in Immunology, 10, 1619.

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